Abstract
Background: Colorectal cancer (CRC) is one of the common tumors of the digestive system. Current CRC therapies have limitations. Huai-Hua-San (HHS) is a traditional Chinese medicine formula comprising two edible herbs, Sophorae Flos and Gardeniae Fructus. HHS was traditionally used to manage TCM symptoms that resemble CRC. However, pharmacological basis of the formula’s application in treating CRC is not known.
Methods: Three CRC cell models were used for in vitro assays, and an HCT116 tumor-bring mouse model was employed for in vivo assays. CCK8 assay was used to examine cell proliferation. Flow cytometry analysis was used to detect apoptosis. Network pharmacology, RNA-Seq and miRNA-Seq were used to predicate signaling pathways involved in the anti-CRC effects of HHS. Immunoblotting was used to examine protein levels, and RT-qPCR was employed to examine mRNA levels.
Results: An HHS extract (HHSE) reduced the viability of, and induced apoptosis in, HCT116, HCT8, and HT-29 CRC cells. Importantly, HHSE dose-dependently inhibited colorectal cancer growth without overt toxicity. KEGG pathway enrichment highlighted the involvement of PI3K/AKT signaling pathway in the anti-CRC effects of HHSE. Western blot results demonstrated that HHSE significantly lowered the protein levels of phospho-AKT (Ser 473). Stimulation with an AKT activator (SC79) significantly reduced the anti-proliferative effects of HHSE, indicating that inhibition of the PI3K/AKT pathway contributes to the anti-CRC effects of the extract. RNA-Seq and miRNA-Seq analyses of mouse tumors suggested that the miR-142-3p/FAM98A pathway was another signal transduction pathway involved in the anti-CRC effects of HHSE. RT-qPCR results showed that HHSE upregulated miR-142-3p levels and downregulated FAM98A mRNA levels in CRC cells. Immunoblotting showed that HHSE significantly downregulated protein levels of FAM98A signaling molecules, including PRMT1, β-catenin, AKT and Bcl-xL.
Conclusions: Our findings provide pharmacological justifications for using HHS in treating CRC and suggest that HHSE can be developed into a modern anti-CRC agent.
Methods: Three CRC cell models were used for in vitro assays, and an HCT116 tumor-bring mouse model was employed for in vivo assays. CCK8 assay was used to examine cell proliferation. Flow cytometry analysis was used to detect apoptosis. Network pharmacology, RNA-Seq and miRNA-Seq were used to predicate signaling pathways involved in the anti-CRC effects of HHS. Immunoblotting was used to examine protein levels, and RT-qPCR was employed to examine mRNA levels.
Results: An HHS extract (HHSE) reduced the viability of, and induced apoptosis in, HCT116, HCT8, and HT-29 CRC cells. Importantly, HHSE dose-dependently inhibited colorectal cancer growth without overt toxicity. KEGG pathway enrichment highlighted the involvement of PI3K/AKT signaling pathway in the anti-CRC effects of HHSE. Western blot results demonstrated that HHSE significantly lowered the protein levels of phospho-AKT (Ser 473). Stimulation with an AKT activator (SC79) significantly reduced the anti-proliferative effects of HHSE, indicating that inhibition of the PI3K/AKT pathway contributes to the anti-CRC effects of the extract. RNA-Seq and miRNA-Seq analyses of mouse tumors suggested that the miR-142-3p/FAM98A pathway was another signal transduction pathway involved in the anti-CRC effects of HHSE. RT-qPCR results showed that HHSE upregulated miR-142-3p levels and downregulated FAM98A mRNA levels in CRC cells. Immunoblotting showed that HHSE significantly downregulated protein levels of FAM98A signaling molecules, including PRMT1, β-catenin, AKT and Bcl-xL.
Conclusions: Our findings provide pharmacological justifications for using HHS in treating CRC and suggest that HHSE can be developed into a modern anti-CRC agent.
| Original language | English |
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| Number of pages | 374 |
| Publication status | Published - 11 Apr 2024 |
| Event | The 7th International Meeting on Intestinal Diseases in conjunction with The Annual Congress of Korean Association for the Study of Intestinal Diseases - Seoul, Korea, Republic of Duration: 11 Apr 2024 → 13 Apr 2024 http://2024.imkasid.org https://imkasid.org/file/IMKASID2024_Program_Book.pdf https://imkasid.org/file/IMKASID2024_Abstract_Book.pdf |
Conference
| Conference | The 7th International Meeting on Intestinal Diseases in conjunction with The Annual Congress of Korean Association for the Study of Intestinal Diseases |
|---|---|
| Abbreviated title | IMKASID 2024 |
| Country/Territory | Korea, Republic of |
| City | Seoul |
| Period | 11/04/24 → 13/04/24 |
| Internet address |
